LOCUS       sequence-470810-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-SRPK1 Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-SRPK1 Vector
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2287..2799)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2838..2856)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2839..2856)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2976..3279
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3434..3454
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3487..3534
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3566..4225
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3632..3651)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4397..4985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4477..4496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5111..5159
                     /note="synthetic polyadenylation signal"
     CDS             complement(5183..5977)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5294..5313)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5904..5923
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6356..7534)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6365..7303
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6388..6408)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 ggagttaagc caagggtgcc ggagacactc ggcggcagtg gctctcttct cagggatcag
      361 ctccaacatg ggcagtaaga aatctgtgaa gccagctgcc tcttcctgcg accactcata
      421 cttctccact agaacctcaa aaaggcccca aggtttcagc ttcgtgatat gtttcaggtc
      481 accttttttg gtgaaaaatt ccttggaata ttttcctgcc acaatgagct tgcgaggcac
      541 cttccccaga agttctatga tcaatgcaat gtgatcttca tctcgagtgt actcttcccc
      601 tgaatgaggt tcaaacaaat agtcacctgt ggccagttca aaggccatgc atgccgtgct
      661 ccaaatgtca gcaggggtat tatagccaga tccgattaga acttccaagg aacgatattg
      721 ccttgtttga atatcttcag tgaaatgttt gtgcacccaa caagcatttc caaggtcagc
      781 aatcttcacc ttgagctttt ctgcattttt tggctcaagg ggattaacaa gaaaatttcc
      841 agccgtggat tttcctttgt tgtccagtgg tccgttatgt tcttgctctt gttcatcttc
      901 acagggtatc tctgcccgaa tgctttcttg aagttggcta atgtgttgtt cactgaatga
      961 ctgacctaca gttgaggaag actggcacac catggtgtct gacacctcag atgttatagg
     1021 tgtacaagag tctgtttctt gagatgtgct gctgtctcca ttttgggagc ttaggaaact
     1081 agattcctga ttcaaatttt ggacatcaca gtcattagca ttatgtagat cctctttatg
     1141 tctcaatgtt tcattattac tgttctgagt ataattaatg acttcaatca ctccattgca
     1201 attaatttct gctgcaccac cctctgtatc acgttccata agcgtttgat cctggccaat
     1261 ggtacttgac tcttcaagtt tttcttgggt cattttatta ggtgggttct ctttcaaggg
     1321 tctttcaaca ggactctctg attcttcttg cttgtttggt cttttttgcc cagggcccga
     1381 ctctttctcc atttcctcaa tttcctgcat tcgcttctct agtaattctg cctggcgctt
     1441 ctgcttcttc ttcaatttct tcttcttatt ctttgacatt ttgtcagctg gtttaggctg
     1501 gggagcagta ctgactgcag atccggaagg cggaggagct ccagatcgct gccattctgt
     1561 tgcttctgca gccagcctcc gaatgtactg ctcattcact gacaataaga tgttctctgg
     1621 tttaatgtca gtgtggatga tacggcactt ggtatgtaaa taatcaagac cctgtaacac
     1681 ttgctgaata atttttttga cacaaggcag tggaagcccc tgataattgg atttgatgat
     1741 ccacttgagc agatgatgcc ccaaaacttc aaataccatg cagatatgtg ttccattaac
     1801 tcctgatatt ttaaagtcat ctagtagttg aacaaccatt tctctatttg gatcattagg
     1861 gtctgaattg cgaactgact tcagcaaccg gatttcatct agtgctgttt cagtgtaatg
     1921 ttcagcactt ttaactactt tcattgccac aaatttcttc ccctgaatat cccatgataa
     1981 ccatactgtt gaaaagtgtc cccagcctaa ctttcggatc acatggtatc tcccattgaa
     2041 tagatctcca attttcacaa gatgataacc tcctttacaa taatcattag gatcttcttg
     2101 ctcatcatca tcagatccca gaatctcctc ttcctgctct ggtagatcac tctcagagtg
     2161 gggagcagag cctcggtgct gagtttcaga tttcctttgg gctttgtcct tcttggcctt
     2221 ggtccttttc tttcgggcct ggagcgcaag cactttccgc tccatggcga tcgcgccgct
     2281 cgagcccgcc agaatgcgtt cgcacagccg ccagccggtc actccgttga tggttactcg
     2341 gaacagcagg gagccgtcgg ggttgatcag gcgctcgtcg ataattttgt tgccgttcca
     2401 cagggtccct gttacagtga tctttttgcc gtcgaacacg gcgatgcctt catacggccg
     2461 tccgaaatag tcgatcatgt tcggcgtaac cccgtcgatt accagtgtgc catagtgcag
     2521 gatcacctta aagtgatgat catccacagg gtacaccacc ttaaaaattt tttcgatctg
     2581 gcccatttgg tcgccgctca gaccttcata cgggatgatg acatggatgt cgatcttcag
     2641 cccattttca ccgctcagga caatcctttg gatcggagtt acggacaccc cgagattctg
     2701 aaacaaactg gacacacctc cctgttcaag gacttggtcc aggttgtagc cggctgtctg
     2761 tcgccagtcc ccaacgaaat cttcgagtgt gaagaccatg gtgagcgcta gctgcaaaaa
     2821 gaacaagtag cttgtattct atagtgtcac ctaaatgggc gatctgacgg ttcactaaac
     2881 cagctctgct tatataggcc tccctggcgc gccactagtt attaatagta atcaattacg
     2941 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3001 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3061 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     3121 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     3181 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     3241 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     3301 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3361 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
     3421 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcaa atcgaattct
     3481 aatactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3541 aggaactaaa cccaggaggc agatcatgga gaaaaaaatc actggatata ccaccgttga
     3601 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     3661 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     3721 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     3781 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     3841 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     3901 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     3961 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     4021 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     4081 tatgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     4141 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     4201 cgatgagtgg cagggcgggg cgtaagcggg actctggggt tcgaaatgac cgaccaagcg
     4261 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4801 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4981 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     5101 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     5161 cggccatcgt gatggctaga gttcagaaga actcgtcaag aaggcgatag aaggcgatgc
     5221 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     5281 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     5341 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     5401 agcaggcatc gccgtgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     5461 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     5521 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     5581 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     5641 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     5701 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     5761 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     5821 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5881 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5941 ccggagaacc tgcgtgcaat ccatcttgtt caatcatggt ggcatcgatt cccggtcaac
     6001 atccagtaca cctccggcac caccggcttc cccaagggcg ccacgctctc ccaccacaac
     6061 atcctcaaca acggctactg ggtgggccgc acggtcggct acacggagca ggaccgggtg
     6121 tgtctgccgg tgcccttcta ccactgcttc ggcatggtca tgggcaactt gggcgccacc
     6181 tcccacgccg cctgcatcgt catcccggcc ccgtccttcg agccggcggc gacgctggag
     6241 gcggtgcagc gggagaggtg cacgtccctg tacggcgtcc ccacgaagcc gaattccacc
     6301 acactggact agatgcatgc tcgacagtgt gatggatatc tgcagaattc ggctttcacg
     6361 acacctgaaa tggaagaaaa aaactttgaa ccactgtctg aggcttgaga atgaaccaag
     6421 atccaaactc aaaaagggca aattccaagg agaattacat caagtgccaa gctggcctaa
     6481 cttcagtctc cacccactca gtgtggggaa actccatcgc ataaaacccc tccccccaac
     6541 ctaaagacga cgtactccaa aagctcgaga actaatcgag gtgcctggac ggcgcccggt
     6601 actccgtgga gtcacatgaa gcgacggctg aggacggaaa ggcccttttc ctttgtgtgg
     6661 gtgactcacc cgcccgctct cccgagcgcc gcgtcctcca ttttgagctg cctgcagcag
     6721 ggccgggaag cggccatctt tccgctcacg caactggtgc cgaccgggcc agccttgccg
     6781 cccagggcgg ggcgatacac ggcggcgcga ggccaggcac cagagcaggc cggccagctt
     6841 gagactaccc ccgtccgatt ctcggtggcc gcgctcgcag gccccgcctc gccgaacatg
     6901 tgcgctggga cgcacgggcc ccgtcgccgc ccgcggcccc aaaaaccgaa ataccagtgt
     6961 gcagatcttg gcccgcattt acaagactat cttgccagaa aaaaagcgtc gcagcaggtc
     7021 atcaaaaatt ttaaatggct agagacttat cgaaagcagc gagacaggcg cgaaggtgcc
     7081 accagattcg cacgcggcgg ccccagcgcc caggccaggc ctcaactcaa gcacgaggcg
     7141 aaggggctcc ttaagcgcaa ggcctcgaac tctcccaccc acttccaacc cgaagctcgg
     7201 gatcaagaat cacgtactgc agccaggtgg aagtaattca aggcacgcaa gggccataac
     7261 ccgtaaagag gccaggcccg cgggaaccac acacggcact tacctgtgtt ctggcggcaa
     7321 acccgttgcg aaaaagaacg ttcacggcga ctactgcact tatatacggt tctcccccac
     7381 cctcgggaaa aaggcggagc cagtacacga catcactttc ccagtttacc ccgcgccacc
     7441 ttctctaggc accggttcaa ttgccgaccc ctccccccaa cttctcgggg actgtgggcg
     7501 atgtgcgctc tgcccactga cgggcaccgg agcc
//
